%0 Journal Article
%T Enhancement of functional expression of wheat peroxidase WP1 in prokaryotic system by co-transforming with hemA and hemL of Esherichia coli<br>共转化大肠杆菌hemA和hemL基因增强小麦过氧化物酶WP1在原核系统中的功能性表达
%A Chao Zhang
%A Liwei Shan
%A Shuaikun Su
%A Yanni Nan
%A Zhongyu Guo
%A Sanhong Fan
%A <br>张超
%A 单丽伟
%A 苏帅坤
%A 南艳妮
%A 郭忠玉
%A 范三红
%J 生物工程学报
%D 2012
%I 
%X Wheat grain peroxidase 1(WP1) belonged to class Ⅲ plant peroxidase with cofactor heme,which not only has antifungal activity,but also influences the processing quality of flour.In order to enhance functional expression of WP1 in prokaryotic system by increasing endogenous heme synthesis,we constructed a recombinant plasmid pACYC-A-L containing hemA and hemL of Esherichia coli.Then,we co-transformed it into host strain T7 Express with secretive expression vector(pMAL-p4x-WP1) or non-secretive expression vector(pET21a-MBP-WP1),respectively.The MBP-WP1 fusion protein was further purified by amylose affinity chromatography and its peroxidase activity was assayed using 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonate)(ABTS) as substrate.At 12 h after induction at 28 degree,the extracellular 5-aminolevulinic acid(5-ALA) production of T7 Express/pACYC-A-L was up to 146.73 mg/L,simultaneously the extracellular porphrins also increased dramatically.The peroxidase activity of functional MBP-WP1 obtained from T7 Express/(pACYC-A-L + pMAL-p4x-WP1) was 14.6-folds of that purified from T7 Express/ pET21a-MBP-WP1.This study not only successfully enhanced functional expression of wheat peroxidase 1 in Esherichia coli,but also provided beneficial references for other important proteins with cofactor heme.
%K wheat peroxidase 1
%K prokaryotic expression
%K hemA
%K hemL
%K heme
%K disulfide bond<br>小麦过氧化物酶1
%K 原核表达
%K 谷氨酰tRNA还原酶
%K 谷氨酸半醛变位酶
%K 血红素
%K 二硫键
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=B39D47F72CB13A7618D9D7807A4EA2EB&yid=99E9153A83D4CB11&vid=D3E34374A0D77D7F&iid=DF92D298D3FF1E6E&sid=461E94ABCF58C63F&eid=BE05E2A2B7E55AA9&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=0